Bioactive packaging

ABSTRACT

Provided herein are packaging materials that include a bioactive material, as well as methods of using and methods of making such bioactive packaging material.

FIELD

Embodiments herein generally relate to bioactive packaging materials,methods of making bioactive packaging materials, and uses of bioactivepackaging materials.

BACKGROUND

There are a variety of packaging materials that can be used to storefood and other materials in order to, for example, preserve and/orprotect the material so stored. Generally, these materials attempt topreserve food by isolating the material from external sources ofcontamination, such as microorganisms.

SUMMARY

In some embodiments, a packaging material is provided. The material caninclude an outer layer that is effectively impermeable to oxygen, aninner layer that is effectively permeable to oxygen, and a bioactivematerial. In some embodiments, the bioactive material is located betweenthe inner layer and the outer layer, and the bioactive layer includes atleast one microorganism.

In some embodiments, a method of containing an item is provided. Themethod can include providing a packaging material that includes an outerlayer that is effectively impermeable to oxygen, an inner layer that ispermeable to oxygen, and a bioactive material. In some embodiments, thebioactive material is located between the inner layer and the outerlayer, and the bioactive layer includes at least one microorganism. Insome embodiments, the method further includes exposing an item to thepackaging material. In some embodiments, a surface of the inner layerfaces a surface of the item to contain the item.

In some embodiments, a method of making a packaging material isprovided. The method can include providing an outer layer that issubstantially impermeable to oxygen, providing a bioactive material, andproviding an inner layer. In some embodiments, the inner layer ispermeable to oxygen, and the bioactive material is positioned betweenthe outer layer and the inner layer, thereby making a packagingmaterial.

The foregoing summary is illustrative only and is not intended to be inany way limiting. In addition to the illustrative aspects, embodiments,and features described above, further aspects, embodiments, and featureswill become apparent by reference to the drawings and the followingdetailed description.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a drawing of some embodiments of a packaging material.

FIG. 2 is a flow chart outlining some embodiments of a method ofcontaining an item.

FIG. 3 is a flow chart outlining some embodiments of a method of makinga packaging material.

DETAILED DESCRIPTION

In the following detailed description, reference is made to theaccompanying drawings, which form a part hereof. In the drawings,similar symbols typically identify similar components, unless contextdictates otherwise. The illustrative embodiments described in thedetailed description, drawings, and claims are not meant to be limiting.Other embodiments may be utilized, and other changes may be made,without departing from the spirit or scope of the subject matterpresented herein. It will be readily understood that the aspects of thepresent disclosure, as generally described herein, and illustrated inthe Figures, can be arranged, substituted, combined, separated, anddesigned in a wide variety of different configurations, all of which areexplicitly contemplated herein.

In some embodiments, a bioactive packaging material is provided. In someembodiments, the bioactive packing material includes one or moremicroorganisms. In some embodiments, the microorganism can be embeddedin a matrix that is part of the packaging material. In some embodiments,the microorganism is initially in a metabolically quiescent state, butcan become activated as an item that is stored in the packaging materialdecomposes. In some embodiments, as the item incubates or decomposes,and/or after the packaging is opened, the amount of oxygen inside thepackaging increases. In some embodiments, the presence of additionaloxygen activates the microorganism, thereby removing it from themetabolically quiescent state. In some embodiments, the microorganismscavenges oxygen, thus decreasing the amount of oxygen in the packagingand preserving the shelf life of the item stored within the packagingmaterial. In some embodiments, the microorganism produces molecules thatprolong the shelf life of the item to be stored. These and otherembodiments are discussed in more detail below.

FIG. 1 is a drawing that illustrates some embodiments of a packagingmaterial 100. In some embodiments, the packaging material includes anouter layer 110. In some embodiments, the packaging material includes aninner layer 120. In some embodiments, the packaging material includes abioactive material 130. In some embodiments, the bioactive material ispart of and/or within the inner layer. In some embodiments, thebioactive material is between the inner layer 120 and the outer layer110 (as depicted in FIG. 1). In some embodiments, the bioactive materialincludes one or more microorganisms 140. While the packaging material isdepicted as a storage container 150, in some embodiments, otherconfigurations and applications are also possible (discussed in moredetail below). In some embodiments, the packaging material has a lid160. In some embodiments, the lid 160 forms an airtight seal with thestorage container 150. In some embodiments, the storage container 150,when an item 101 is placed in it, includes a headspace 170.

A variety of outer layers can be employed. In some embodiments, theouter layer is effectively impermeable to liquid. In some embodiments,the outer layer effectively prevents 50, 60, 70, 80, 90, 95, 96, 97, 98,99, 99.9, 99.99, 99.999% or more of a liquid from passing through it. Insome embodiments, the outer layer is effectively impermeable to gas. Insome embodiments, the outer layer effectively prevents 10, 20, 30, 40,50, 60, 70, 80, 90, 95, 96, 97, 98, 99, 99.9, 99.99, 99.999, 99.9999,99.99999% or more of a gas from passing through it. In some embodiments,the gas is oxygen. In some embodiments, the outer layer limits diffusionof oxygen. In some embodiments, the outer layer limits diffusion ofoxygen across it so that at a temperature range of −50° C. to 150° C.,there is a first partial pressure of oxygen on one side of the outerlayer, which is different from a second the partial pressure of oxygenon the other side of the outer layer, and after 24 hours have passed, atleast about 80% of that difference in partial pressure of oxygen stillexists. In some embodiments, the outer layer effectively prevents anamount of oxygen from passing through it that would be enough toactivate the microorganisms in the packaging material. In someembodiments, the outer layer provides an effectively airtight barrierover the period of time for which the packaging material is to be usedto store an item (which can be, for example, hours, days, weeks, months,or years).

In some embodiments, the outer layer is at least about 0.001 millimetersthick, e.g., 0.001, 0.002, 0.005, 0.01, 0.03, 0.05, 0.10, 0.15, 0.2,0.5, 1, 2, 3, 5, 10, 15, 20, 30, 50, 80, 100, or 200 millimeters thick,including any range defined between any two of the stated values and anyrange above any one of the stated values.

In some embodiments, the outer layer includes a polymer. In someembodiments, the outer layer is biodegradable. In some embodiments, theouter layer can include plastic, metal, and/or glass.

In some embodiments, the packaging material includes an inner layer;however, the inner layer can be optional. In some embodiments, the innerlayer is effectively permeable to gas. In some embodiments, the gas isoxygen, carbon dioxide, or both oxygen and carbon dioxide. In someembodiments, the inner layer is permeable to one or more of thefollowing: an antimicrobial compound, a preservative, a flavor modifier,an odor molecule. In some embodiments, the inner layer is simply partof, or is, a matrix, which can contain the microorganisms. In someembodiments, the inner layer includes a polymer. In some embodiments,the inner layer includes a film. In some embodiments, the inner layer isa polymer film. In some embodiments, the polymer film includes ahomopolymer. In some embodiments, the polymer includes a heteropolymer.In some embodiments, polymer film includes a plastic polymer and/or acellulose polymer. In some embodiments the inner layer includes aninsert, for example a plastic, cardboard, various polymers, PVDF, waxes,lipids, silicates rubber, and/or cellulose. In some embodiments, theinsert contacts the matrix. In some embodiments, the insert iscovalently bonded to at least a part of the matrix. In some embodiments,the insert is positioned further than 0.5 micrometers from the matrix,e.g., 1, 2, 3, 5, 10, 15, 20, 30, 50, 80, 100, 150, 200, 250, 300, 400,500, 700, 900, 1000, 1500, 2000, 3000, or 5000 micrometers from thematrix, but does not contact the matrix (including any range above anyone of the preceding values and any range defined between any two of thepreceding values).

In some embodiments, the inner layer has a thickness of at least about0.5 micrometers, e.g., 0.5, 1, 2, 3, 5, 10, 15, 20, 30, 50, 80, 100,150, 200, 250, 300, 400, 500, 700, 900, 1000, 1500, 2000, 3000, 5,000,10,000, or 100,000 micrometers, including any range above any one of thepreceding values and any range defined between any two of the precedingvalues. In some embodiments, the inner layer is opaque. In someembodiments, the inner layer is transparent. In some embodiments, theinner layer contains perforations or pores. In some embodiments theperforations or pores have diameters of at least about 0.1 micrometer,e.g., 0.1, 0.3, 0.7, 1, 2, 3, 5, 10, 15, 20, 30, 50, 80, 100, 150, 200,250, 300, 400, 500, 700, 900, 1000, 1500, 2000, or 3000 micrometers,including any range above any one of the preceding values and any rangedefined between any two of the preceding values. In some embodiments,the perforations or pores have substantially uniform diameters. In someembodiments, the perforations or pores have diameters of two or moresubstantially different sizes.

In some embodiments, the packaging material includes a matrix, which, insome embodiments, can be the bioactive layer. In some embodiments, themicroorganism contacts and/or is associated with the matrix. In someembodiments, the microorganism is embedded in the matrix. In someembodiments, the microorganism contacts a surface of the matrix. In someembodiments, the matrix is located between the inner layer and the outerlayer. In some embodiments, the matrix is, or is part of, the innerlayer. In some embodiments, there is no inner layer, and the matrix isexposed to the interior of the packaging material. In some embodiments,the matrix contacts the item or items to be packaged by the packagingmaterial. In some embodiments, the matrix is permeable to gas. In someembodiments, the matrix is permeable to oxygen. In some embodiments, thematrix is permeable to carbon dioxide. In some embodiments, the matrixis a solid. In some embodiments, the matrix is a foam. In someembodiments, the matrix is a gel.

In some embodiments, the matrix includes a nutrient for themicroorganism. In some embodiments, the matrix includes at least one ofa starch, a carbohydrate, a polynucleotide, a protein, or a syntheticpolymer. In some embodiments, these are included in an amount sufficientfor keeping the microorganism alive for a period of time during whichthe packaging material is to be used, e.g., at least hours, days, weeks,months, or years.

In some embodiments, the matrix includes a cross-linking agent. In someembodiments, molecules of the matrix are cross-linked. In someembodiments, the matrix includes at least one of a peptide, a caseinpeptone, a vitamin, for example a B vitamin, nitrogen, sulfur,magnesium, or a mineral, for example an essential mineral.

In some embodiments, one or more of the components of the matrix isbiodegradable. In some embodiments, all or substantially of theingredients of the matrix are biodegradable.

In some embodiments, the matrix does not include a bioactive material,such as a microorganism. For example, in some embodiments, an outerlayer with a matrix applied to the inside of the outer layer can beprovided as a product. At a later point in time, one can add one or moremicroorganisms to the matrix, allow them to attach to the matrix, andthen use the packaging material. Thus, in some embodiments, any of thepackaging material embodiments provided can be provided as an “inert”form (without the microorganism), and the microorganism can be addedlater (e.g., directly to the matrix, to the matrix and crosslinked,directly to an internal surface of the outer layer, directly to theouter surface of an inner layer, etc.) In some embodiments, these itemscan be provided in the form of a kit.

In some embodiments, the packaging material includes a bioactivematerial 130. In some embodiments, the bioactive material 130 isseparate from the inner layer 120. In some embodiments, the bioactivematerial 130 is located between the outer layer 110 and the inner layer120. In some embodiments, the bioactive material 130 is part of theinner layer (e.g., the bioactive material can be located within and/oron the inner layer). In some embodiments, there is no inner layer, andthe bioactive material is exposed to the interior of the packagingmaterial. In some embodiments, there need not be an inner layer and thebioactive material can be exposed to the interior of the packagingmaterial. In such embodiments, the inner surface of that layer can be,for example, crosslinked. In some embodiments, the bioactive materialcan be included within a matrix. In some embodiments, the bioactivematerial includes one or more microorganism, e.g., at least 1, 10, 100,1,000, 10,000, 100,000, 1,000,000, 10,000,000, 100,000,000,1,000,000,000, 10,000,000,000, 100,000,000,000, or more microorganisms.In some embodiments, the number of microorganisms per cubic centimetercan be determined based upon the particular application. In someembodiments, the number of microorganisms per cubic centimeter can befrom 1 to 1*10̂11 cells/mL.

In some embodiments, the microorganism is in a metabolically quiescentstate. In some embodiments, the metabolically quiescent state is a “livebut not replicating (LNR)” state. In some embodiments, the LNR stateprovides reduced replication by the microorganism. In some embodiments,the LNR state includes a replication rate that is no faster than about1×10⁻¹, 5×10⁻¹, 1×10⁻², 1×10⁻³, 1×10⁻⁴, 1×10⁻⁵, or 1×10⁻⁶ of thereplication rate of the microorganism cultivated on rich medium in anormal, non-metabolically quiescent state. In some embodiments thereplication rate is effectively zero. In some embodiments, themetabolically quiescent state includes a state of nutrient deprivation.In some embodiments, the metabolically quiescent state is “metabolichibernation.” In some embodiments, the metabolically quiescent state isreversible so that the microorganism will exit the metabolicallyquiescent state and return to metabolically active state. In someembodiments, a substance secreted by the packaged item induces themicroorganism to exit the metabolically quiescent state. In someembodiments, oxygen induces the microorganism to exit the metabolicallyquiescent state. In some embodiments, the microorganism is geneticallymodified to enter a quiescent state. For example, in some embodimentsthe microorganism contains a mutation in a regulator of the cell cycleor metabolic enzyme that reduces the growth rate of the bacterium.Furthermore, in some embodiments, the microorganism optionally includesan inducible transgene that restores normal cell cycle regulation ormetabolism in the presence of a desired molecule, for example oxygen. Insome embodiments, a Tet ON/OFF system can be used. For example, in thepresence of Tetracycline (for example in a Tet OFF system), thetransgene of interest is repressed. This can be used, to switch themicroorganisms in the matrix “on” when the tetracycline in the food isdepleted and/or destroyed. This can be used, for example, as a secondline antibacterial defense by having the microorganisms in the packagingproduce bacteriocins in the absence of the antibiotic.

In some embodiments, the microorganism is aerobic. In some embodiments,the microorganism is aerobic, and the metabolically quiescent stateincludes oxygen deprivation. In some embodiments, the microorganismscavenges oxygen. In some embodiments, the microorganism secretes one ormore substances. In some embodiments, the microorganism scavenges oxygenand secretes one or more substances.

In some embodiments the microorganism is prokaryotic. In someembodiments, the microorganism is a bacterium. In some embodiments, themicroorganism is eukaryotic. In some embodiments, the microorganism is afungus. In some embodiments, the microorganism is a yeast. In someembodiments, a mixture of microorganisms of two or more types is used.In some embodiments, the microorganism is one that is GenerallyRecognized As Safe (GRAS) in the United States, Australia, Europe,and/or Japan. In some embodiments, the microorganism can include one ormore of Lactic acid bacteria (LAB), bifidobacteria, yeast, bacilli,Bacillus coagulans, Lactobacillus acidophilus, Lactobacillus casei,Lactobacillus casei, Shirota, non-pathogenic microorganisms, bacteria,Acetobacter, Acholeplasma laidlawii, Acidovorax, Actinoalloteichus,Actinosynnema minim, Aeromicrobium, Agrobacterium, tumefaciens,Lactobacillus casei, Alicyclobacillus, Alishewanella, Aneurinibacillus,Aquabacterium commune, Aquabacterium citratiphilum, Aquabacteriumparvum, Aquaspirillum itersonii, Aquifex aeolicus, Aquifex pyrophilus,Arthrobacter globiformis, Azomonas macrocytogenes, Bacillus agri,Bacillus alginolyticus, Bacillus aneurinolyticus, Bacillus azotoformans,Bacillus atrophaeus, Bacillus badius, Bacillus borstelensis, Bacilluscentrosporus, Bacillus chondroitinus, Bacillus choshinensis, Bacilluscirculans, Bacillus coagulans, Bacillus cohnii, Bacillus formosus,Bacillus galactophilus, Bacillus globisporus, Bacillus halodurans,Bacillus laevolacticus, Bacillus licheniformis, Bacillus megaterium,Bacillus methanolicus, Bacillus migulanus, Bacillus mojavensis, Bacillusmycoides, Bacillus naganoensis, Bacillus pallidus, Bacillus parabrevis,Bacillus polymyxa, Bacillus pumilus, Bacillus reuszeri, Bacillussphericus, Bacillus stearothermophilus, Bacillus subtilis, Bacillusthermocloacae, Bacillus thuringiensis, Blastomonas, Brachybacterium,Brochothrix, Brevibacillus, Brevibacterium, Brevundimonas vesicularis,Budvicia aquatica, Buttiauxella agrestis, Butyrivibrio crossotus,Carnobacterium, Carnobacterium divergens, Caulobacter, Cellulomonas,Cellulomonas cellulans, Clostridium butyricum, Clostridium tertium,Clostridium tetanomorphum, Collinsella intestinalis, Collinsella spp,Collinsella stercoris, Comomonas acidovorans, Corynebacterium accolens,Corynebacterium afermentans, Corynebacterium argentoratense,Corynebacterium auris, Corynebacterium genitalium, Corynebacteriumpropinquum, Corynebacterium pseudodiphtheriticum, Corynebacteriummacginleyi, Corynebacterium tuberculostearicum, Corynebacteriumurealyticum, Curtobacterium, Deinococcus spp, Delftia, Dermacoccusnishinomiyaensis, Desemzia, Dietzia, Dysgonomonas, Empedobacter,Enterobacter agglomerans, Enterococcus avium, Enterococcus durans,Enterococcus porcinus, Enterococcus ratti, Erwinia, Escherichia blattae,Exiguobacterium, Fibrobacter, Filifactor, Finegoldia, Flavobacteriumcapsulatum, Flavobacterium columnare, Flavobacterium psychrophilum,Fusobacterium prausnitzii, Geotrichium candidum, Gluconobacter,Glycomyces tenuis, Gracilibacillus, Granulicatella, Haemophilusparagallinarum, Haemophilus parasuis, Haemophilus somnus, Halobacteriumsalinarium, Helicobacter hepaticus, Helicobacter muridarum, Holdemania,Intrasporangium calvum, Klebsiella terrigena, Kocuria, Kocuria rosea,Kurthia gibsonii, Kytococcus, Lactobacillus delbrueckii, Lactobacillusfermentum, Lactobacillus leichmannii, Lactobacillus oris, Lactobacillusplantarum, Lactobacillus vaginalis, Lactococcus garvieae, Lactococcuslacti, Lautropia, Lawsonia, Lechevaliera, Leifsonia, Lentzia, Leptospirabiflexa, Leuconostoc, Listeria innocua, Listeria ivanovii, Listeriawelshimeri, Luteococcus, Macrococcus, Mannheimia, Maricaulis, Megamonas,Methylobacterium amnivorans, Methylobacterium mesophilicum, Micrococcusdiversus, Mycoplasma gallisepticum, Micrococcus luteus, Micrococcusroseus, Micromonas, Micromonospora coerulea, Moraxella bovis,Mortierella wolfii, Mucor hiemalis, Mycoplasma bovigenitalium,Mycoplasma hyopneumoniae, Mycoplasma hyorhinis, Mycoplasma iowae,Mycoplasma synoviae, Mycoplasma orale, Nesterenkonia, Obesumbacteriumproteus, Oerskovia, Oligella ureolytica, Ornithobacterium,Paenibacillus, Paracoccus, Penicillium expansum, Penicillium verrucosum,Pentatrichomonas hominis, Planobispora rosea, Porphyromonasendodontalis, Porphyromonas gulae, Pragia fontium, Propioniferax,Proteus myxofaciens, Pseudomonas alcaligenes, Pseudomonas diminuta,Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas syringae,Renibacterium salmoninarum, Ruminococcus, Rhodospirillum rubrum,Rickenella, Ruminococcus productus, Saccharothrix longispora,Saccharothrix mutabilis, Sanguibacter, Schineria, Sebaldella, Shewanellaputrefaciens, Slackia, Solobacterium, Sporosarcina ureae, Staphyoloccuscaprae, Staphylococcus carnosus, Staphylococcus lentus, Staphylococcuspulvereri, Stomatococcus, Streptococcus parauberis, Streptomyces albus,Streptomyces corchorusii, Streptomyces olivaceoviridis, Streptomycesscabiei, Streptosporangium roseum, Taylorella, Tatlockia,Tetragenococcus halophilus, Terracoccus, Thermoanaerobacteriumthermosaccharolyticum, Thermotoga maritime, Thermus, Tissaracoccus,Tritrichomonas foetus, Turicella otitidis, Ureaplasma diversum,Vagococcus fluvialis, Vagococcus salmoninarum, Xanthomonas campestris,Xenorhabdus nematophilus, Yersinia ruckeri, Zoogloea ramigera,Zygosaccharomyces rouxii, Fungi, Acremonium strictum, Aspergilluspenicillioides, Dactylaria gallopava, Galactomyces geotrichum,Metschnikowia pulcherrima, Neurospora, Penicillium notatum, Penicilliumroquefortii, Pentatrichomonas hominis, Pichia haplophila, Saccharomycescarlsbergensis, Saccharomyces pastorianus, Schizosaccharomyces,Tritrichomonas mobilensis, Zygosaccharomyces bailii, and/orZygosaccharomyces rouxii. In some embodiments, the organism can be onethat is an aerobe that is GRAS, such as Bacillus subtillis, that canferment under some conditions.

In some embodiments, the microorganism is a probiotic organism that isused as a nutritional supplement. In some embodiments, the number oforganisms used, as well as the concentration in the final packagingproduct will depend on the type of organism chosen, as each can have asomewhat different respiration rate.

In some embodiments, the microorganism can survive at a temperature ofat least 0° C., for example 10, 10, 20, 30, 50, 60, 70, 80, 90, 100,110, 120, 140, 170, 200, or 250° C., including ranges between any two ofthe listed values and any range above any one of the preceding values.In some embodiments, the microorganism can survive at a pressure of atleast 100 kilopascals, e.g., 100, 110, 120, 130, 140, 150, 160, 170,180, 190, 200, 240, 300, 350, 400, or 500 kilopascals, including rangesbetween any two of the listed values and any range above any one of thepreceding values. In some embodiments, the microorganism can survive athigh temperature (above room temperature) and high pressure (aboveatmospheric). In some embodiments the microorganism is a thermophile. Insome embodiments, the microorganism is has undergone selection, and/oris genetically modified to facilitate survival at extreme temperatureand/or pressure.

In some embodiments, the microorganism secretes one or more substances.In some embodiments, the microorganism is genetically modified tosecrete a substance. In some embodiments, the substance includes aflavor modifier, color modifier, or odor modifier. In some embodiments,the substance can be a pH modifier (for example, an acid or a base),antimicrobial (for example, bacteriocin), texture modifiers (forexample, gelatin, proteins, polysaccharide) sweetener (for example,sugar), etc. In some embodiments, the microorganism secretes thesubstance in response to a condition, for example high oxygenconcentration or spoilage of a packaged item. In some embodiments, thesecreted substance warns a user, manufacturer, or vendor of a conditionof the item. In some embodiments, the secreted substance provides avisual cue. In some embodiments, the visual cue can be observed throughthe packaging, and thus, one need not open the packaging to observe thecue, when, for example, the packaging is at least partially transparent.In some embodiments, the compounds could include luciferase/luciferin,and/or could be a pigment produced by the micoorganisms, such asmelanin, or another pigmented protein.

In some embodiments, the packaging material is opaque. In someembodiments, the packaging material is effectively transparent. In someembodiments, the packaging material is at least about 30%, 50%, 70%,80%, 90%, 95%, or 99% transparent to visible light, including rangesbetween any two of the stated values. In some embodiments, at least onelayer of the packaging material is opaque, and at least one layer of thepackaging material is transparent. For example, in some embodiments thematrix and/or inner layer is opaque, while the outer layer iseffectively transparent to permit visual examination of the matrixand/or bioactive material.

In some embodiments, the outer layer, the inner layer, and/or the matrixprovides a rigid wall of the packaging material. In some embodiments,all three of the outer layer, the inner layer, and the matrix togetherprovide a rigid wall. In some embodiments, the outer layer and innerlayer provide a rigid wall. In some embodiments, the outer layer andmatrix provide a rigid wall. In some embodiments, the inner layer andmatrix provide a rigid wall. In some embodiments, at least one of theouter layer, inner layer, or matrix provides a flexible wall, forexample a conforming wall that conforms to the item being packaged. Insome embodiments, the inner layer and/or matrix provide a conformingwall while the outer layer is rigid. In some embodiments, all three ofthe inner layer, matrix, and outer layer conform to the item beingpackaged.

In some embodiments, the packaging material is in the shape of acontainer. In some embodiments, the packaging material is in the shapeof a free standing storage container. In some embodiments, the storagecontainer is one of a canister, a box, a jar, a bottle, a capsule, aspecimen tube, a blister pack, a pouch, a bag, etc.

In some embodiments, the storage container is a food storage container.In some embodiments, the storage container is an agricultural productstorage container, for example, for storing plants, seeds, and/orspores. In some embodiments, the storage container is a biologicalspecimen container. In some embodiments, the storage container is a cellculture container. In some embodiments, the storage container is anorgan transport container. In some embodiments, the storage containerstores a forensic sample. In some embodiments, the storage container canbe (and/or configured) for pharmaceuticals, biologics, film,electronics, paint, various coatings, and/or fast moving consumer goods(FMCG).

In some embodiments, the storage container has an opening. In someembodiments, the storage container has two or more openings. In someembodiments the container is, and/or can be sealed, such that it issubstantially airtight unless an opening in the material is provide oris opened. In some embodiments, the opening is configured to becloseable and can include with an airtight seal. In some embodiments,the opening is for inserting or removing the item to be packaged. Insome embodiments, the opening is a ventilation opening. In someembodiments, the opening has a diameter of about at least about 0.1 cm,e.g., 0.1, 0.5, 1, 2, 3, 5, 8, 10, 12, 15, 20, 25, 30, 40, 50, 60, 80,100, 150, 200, 250, 300, or 500 cm, included any range between any twoof the listed values and any range above any one of the precedingvalues. In some embodiments the seal is reusable, and the container canundergo multiple cycle of unsealing and re-sealing. In some embodiments,the seal is a single use seal.

In some embodiments, the container includes a lid. In some embodiments,the lid includes a gasket. In some embodiments, the lid forms anairtight seal with the container.

In some embodiments, for example when the packaging material is rigid,the storage container seals a substantially fixed volume, which can bedefined by the packaging material. In some embodiments, for example whenthe packaging material is flexible or conforming, the storage containerseals a maximum volume, which can be defined by the packaging material.In some embodiments, when the storage container is sealed, it encloses avolume of at least about 0.00001 liters, e.g., 0.00001, 0.0001, 0.001,0.01, 0.1, 0.3, 0.5, 0.7, 1, 1.3, 1.5, 1.8, 2, 2.5, 3, 4, 5, 7, 10, 15,20, 50, 100, 200, 300, 500, 1000, 10,000, or 100,000 liters, includingranges between any two of the listed values and any range above any oneof the preceding values. In some embodiments, the storage containerincludes two or more compartments and when the container is sealed, eachcompartment is substantially impermeable to gas.

In some embodiments, the opening has a self-sealing closure. In someembodiments, the self-sealing closure is resealable. In someembodiments, the self-sealing closure can be opened and resealed atleast 1 time, e.g., about 1, 2, 3, 5, 10, 15, 20, 40, 50, 100, 150, 200,300, or 500 times, including any range between any two of the precedingvalues. In some embodiments, the self-sealing closure includes anadhesive. In some embodiments, the adhesive is pressure sensitive. Insome embodiments, the self-sealing closure includes a zipper. In someembodiments, the self-sealing closure includes a clasp. In someembodiments, the storage container is a self-sealing bag.

In some embodiments, a method of containing an item is provided. In someembodiments, one or more item are contained, e.g., 1, 2, 5, 10, 100,1000, 10,000, 100,000 or more item can be contained within the packagingmaterial. In some embodiments, the method includes providing a packagingmaterial that includes a bioactive material and exposing the item to thepackaging material so that an inner surface of the packaging materialfaces the item.

FIG. 2 is a flow chart that illustrates some embodiments of a method ofcontaining an item 200. In some embodiments, a packaging material thatincludes an outer layer that is effectively impermeable to gas isprovided, along with a bioactive material and (optionally) an innerlayer that is effectively permeable to gas 210. In some embodiments, theitem is exposed to the packaging material so that an inner surface ofthe packaging material faces a surface of the item 220. Optionally, insome embodiments, the item can be fully contained within the packagingmaterial 230 for some period of time thereafter.

One skilled in the art will appreciate that, for this and otherprocesses and methods disclosed herein, the functions performed in theprocesses and methods may be implemented in differing order.Furthermore, the outlined steps and operations are only provided asexamples, and some of the steps and operations may be optional, combinedinto fewer steps and operations, or expanded into additional steps andoperations without detracting from the essence of the disclosedembodiments.

In some embodiments, the method of containing an item includes providingany of the packaging material embodiments described herein. In someembodiments, the method includes exposing the item to the packagingmaterial, so that an inner surface of the packaging material faces asurface of the item. In some embodiments, a surface of the item is incontact with an inner surface of the packaging material. In someembodiments, the item is not in contact with an inner surface of thepackaging material, but gas can diffuse between the item and the innersurface of the packaging material. In some embodiments, the methodincludes sealing the item within the packaging material. In someembodiments, the method includes wrapping the item in the packagingmaterial. In some embodiments, the method includes spraying thepackaging material onto the item (e.g., each layer can be spray appliedonto the item, finishing with the outer layer).

In some embodiments, the method includes positioning the item to bepackaged proximal to the inner layer, and positioning the item to bepackaged distal to the outer layer. In some embodiments, the methodincludes positioning the item to be packaged proximally to the bioactivematerial, and positioning the outer layer distal to the item to bepackaged.

In some embodiments, the item to be packaged is a food product. In someembodiments, the item to be packaged is one of a fruit, a vegetable, adairy product, a meat product, a baked product, a ready-to-eat item, abeverage, or a combination of the above. In some embodiments, the itemto be packaged is an agricultural product, for example a plant, seed,spore, or culture. In some embodiments, the item to be packaged is acell culture. In some embodiments, the item to be packaged is an organor tissue, for example for transplant or laboratory analysis. In someembodiments, the item to be packaged is a forensic sample. In someembodiments, the item to be packaged is a pharmaceutical product or anyof the other products and/or materials described herein.

In some embodiments the packaging material is (and/or is part of) acontainer. In some embodiments, the method includes inserting the iteminto a container that includes the packaging material. In someembodiments, the method includes assembling the container around theitem to be packaged.

In some embodiments, the method includes sealing the container. In someembodiments, the item to be packaged is sealed inside of the container.In some embodiments, sealing includes applying pressure to apressure-sensitive adhesive. In some embodiments, sealing includesclosing a zipper closure. In some embodiments, sealing includes closinga clasp. In some embodiments, sealing includes applying a lid, andsecuring the lid, for example with friction, with a screw, with a clasp,with a strap, with a sealant, and/or with a tape. In some embodiments,sealing includes applying a substantially airtight seal. In someembodiments, the lid, zipper closure, and/or adhesive closure forms anairtight seal when it contacts the container. In some embodiments,applying the airtight seal includes contacting the lid or closure with agasket. In some embodiments, applying the airtight seal includesapplying a sealant, for example a gel, putty, foam, paste, tape, and/orfilm. In some embodiments, the sealant includes an adhesive. In someembodiments, after sealing, the container is reopened and then resealed.In some embodiments, multiple cycles of opening and re-sealing areperformed.

In some embodiments, the method includes sealing the container so thatthere is a headspace volume between the item to be packaged and theinner surface of the container (see FIG. 1, 170). In some embodiments,gas effectively flows between the item and the headspace. In someembodiments, the headspace volume is at least about 0.1 percent of thevolume of the container, e.g., 0.1, 0.2, 0.3, 0.5, 1, 1.5, 2, 3, 5, 10,15, 20, 30, 50, 60, 70, 80, 90, 95, 99, 99.9 percent, including rangesbetween any two of the listed values and any range below any one of thepreceding values.

In some embodiments, immediately after the item is sealed in thecontainer, the headspace volume contains some amount, e.g., a particularpercentage content, of oxygen. In some embodiments, 2 hours after theitem is sealed in the container, the percentage content of oxygen in theheadspace decreases, and/or the percent content of carbon-dioxideincreases. In some embodiments, the percentage content of oxygendecreases at least about 1%, 3%, 5%, 10%, 20%, 30%, 50%, 60%, 70%, 80%,90%, 95%, 98%, 99% or more in comparison to the amount present at thetime the item was sealed in the container, including ranges between anytwo of the listed values. In some embodiments, the percentage content ofcarbon dioxide increases at least about 1%, 3%, 5%, 10%, 20%, 30%, 50%,60%, 70%, 80%, 90%, 95%, 98%, 99% or more in comparison to the amountpresent at the time the item was sealed in the container, includingranges between any two of the listed values.

In some embodiments, the method allows gas to travel between theheadspace 170 to the bioactive material 130 (FIG. 1). In someembodiments, this occurs by gas diffusing between the headspace and thebioactive material. In some embodiments, for example when the innerlayer is present, gas travels through the inner layer to arrive at thebioactive material, for example by diffusion. In some embodiments, thegas includes oxygen. In some embodiments, the microorganism in thebioactive material is in a metabolically quiescent state prior to beingcontacted with oxygen, and contact with oxygen induces the microorganismto enter a metabolically active state, thereby allowing themicroorganism to convert oxygen to CO₂.

In some embodiments, microorganism of the bioactive material metabolizesthe oxygen that is present in the container and the microorganismexchanges the oxygen for carbon dioxide. In some embodiments, the carbondioxide travels from the bioactive material to the headspace. In someembodiments, for example when the inner layer is present, the carbondioxide travels through the inner layer, for example by diffusion. Thus,the exchange by the microorganisms of oxygen for carbon dioxide reducesthe percentage content of oxygen in the headspace. In some embodiments,the microorganism metabolizes oxygen, and incorporate the oxygen into anon-elemental oxygen molecule, for example water.

In some embodiments, any of the above methods can be used for increasingand/or preserving various items, such as food, pharmaceutical products,etc. In some embodiments, any of the above methods do not need toincrease the shelf life of an item, but instead (or in addition to) canprovide a method for providing a flavor modifier, a color modifier,and/or odor modifier. In some embodiments, any of the above methods donot need to increase the shelf life of an item, but instead (or inaddition to) can provide a method for warning a user, manufacturer,and/or vendor of a condition of the item. In some embodiments, asecreted substance from the bioactive material provides a visual cue. Insome embodiments, the visual cue can be observed through the packaging,and thus, one need not open the packaging to observe the cue, when, forexample, the packaging is at least partially transparent.

In some embodiments, a method of making a packaging material isprovided. In some embodiments, the method includes providing a bioactivematerial. In some embodiments, the method includes providing an outerlayer that is substantially impermeable to gas and associating thebioactive material with the outer layer, either directly or indirectly.

FIG. 3 is a flow chart that depicts some embodiments of a method ofmaking a packaging material. In some embodiments, an outer layer that iseffectively impermeable to gas is provided 300 and a bioactive materialcan then be provided 310 and associated with the outer layer. In someembodiments, an inner layer that is effectively permeable to gas can,optionally, be provided 320. In some embodiments, the inner layer ispositioned so as to keep various items in the container or packagingmaterial separated from the bioactive material, but to still allow gasand/or other substances to pass to and from the bioactive material. Insome embodiments, the bioactive material can be associated with an innersurface of the outer layer, or another layer associated with the outerlayer 330. In some embodiments, the bioactive layer is placed directlyonto the inner surface of the outer layer. In some embodiments, thebioactive layer is placed directly on the inner layer, and the innerlayer is placed adjacent to, and therefore associated with, the innersurface of the outer layer. In some embodiments, the outer layer is partof a container, and the bioactive material is positioned on an innersurface of the outer layer. In some embodiments, one or more additionallayers can be provided between the bioactive layer and the outer layerand/or the bioactive layer and the inner layer. In some embodiments, thebioactive material is positioned on a surface of the inner layer. Insome embodiments, the inner layer is part of a container, and thebioactive material is positioned on an outer surface of the inner layer.

In some embodiments, the microorganism can be cultured in ametabolically normal state and then induced into a metabolicallyquiescent state. In some embodiments, the batch is purified beforeand/or after induction into the metabolically quiescent state. In someembodiments, the microorganism is induced into the metabolicallyquiescent state through at least one of: nutrient deprivation,incubation at a temperature substantially different from the normalculturing temperature of the microorganism, incubation of the microbeunder osmotic conditions substantially different from normal osmoticconditions, oxygen deprivation (for example, for aerobicmicroorganisms), contacting the microorganism with a food preservative,and/or genetically modifying the microorganism. In some embodiments, thenutrients deprived from the microorganism include one or more ofcarbohydrates, nitrogen, and essential amino acids. In some embodiments,the microorganism is cultivated in a hypertonic solution.

In some embodiments, the microorganism can be embedded in a matrix. Insome embodiments, embedding the microorganism in the matrix includescombining the microorganism with a matrix material. In some embodiments,the microorganism is mixed with at least one ingredient that can be usedto form the matrix. In some embodiments, the microorganism is added to amixture that includes monomers and/or subunits for the creation of amatrix polymer. In some embodiments, embedding the microorganism in thematrix includes polymerizing the monomers and/or subunits such that themicroorganism is trapped and/or associated with the matrix materialafter the polymerization. In some embodiments, embedding themicroorganism in the matrix includes cross-linking the matrix material.In some embodiments, embedding the microorganism in the matrix includespolymerizing and cross-linking.

In some embodiments, the matrix material is applied to a layer of thepackaging material. In some embodiments, the matrix is applied to alayer of the packaging material, and the layer is then incorporated intoa container. In some embodiments, a layer of the packaging material isincorporated into a container and the matrix is applied to the layer. Insome embodiments, the matrix material is applied to an inner surface ofthe outer layer. In some embodiments, the matrix material is applied toan outer surface of the inner layer. In some embodiments, two layers ofthe packaging material are incorporated into a container, and the matrixis inserted between the layers, for example between the inner layer andthe outer layer. In some embodiments, the matrix is sprayed onto one orboth layers. In some embodiments, the matrix is spread onto the layer.In some embodiments, the matrix is poured onto a layer. In someembodiments, the matrix material is spread, sprayed, or poured onto alayer and then cross-linked, polymerized, or polymerized andcross-linked to increase its viscosity and/or trap and/or associate amicroorganism with the matrix material.

In some embodiments, the microorganism is embedded in the matrix beforethe matrix is applied to the layer, for example by mixing themicroorganism with matrix ingredients. In some embodiments, themicroorganism is added to the matrix after the matrix is applied to thelayer, for example by spreading the microorganism, or applying themicroorganism via an aerosolized spray. In some embodiments, themicroorganism is applied to the matrix material after the matrixmaterial is crosslinked and/or polymerized.

In some embodiments, the matrix can be used as a skeleton for thepackaging material and the outer layer can be sprayed onto the outsideof the material, the microorganism can be added to the matrix, and thenthe inner layer can be sprayed inside the packaging material. In someembodiments, there need be no inner layer, if, for example, themicroorganisms are harmless to the material to be stored and/or themicroorganisms are fixed within a matrix or to the outer layer.

In some embodiments, the matrix includes the microorganisms, togetherwith a matrix, and some basic nutrients and salts, in order to maintainviability, such as one or more of: peptides and casein peptones,vitamins (including B vitamin), trace elements (e.g. nitrogen, sulfur &magnesium), and/or minerals.

In some embodiments, the matrix can be made from a variety of materials,such as cross-linked starches, other carbohydrates, polynucleotides,proteins, or synthetic polymers. In some embodiments, the matrix andmicroorganisms are placed under high pressure, for example about 120kilopascals, 150, 170, 190, 200, 210, 230, 250, 300, 400, or 500kilopascals to facilitate spraying to form the matrix. In some cases,this is sprayed to the inner barrier, and then applied (e.g. as aninsert) to the outer barrier. In some cases, the matrix is cross-linkedafter application, such as by adding a cross-linking agent whichspecifically bonds the matrix polymers together, or specificallypolymerizes the matrix material. In some cases, this is accomplished byheating and/or drying the matrix material, although the viability of themicroorganisms needs to be taken into account with these methods.

In some embodiments, the microorganism is associated directly to atleast one of the inner layer or the outer layer. In some embodiments,this can be achieved by hydrophobic forces. In some embodiments, thiscan be achieved by cross-linking or other covalent bonds.

In some embodiments, the packaging materials provided herein can involvecomponents that are recyclable and/or biodegradable.

EXAMPLES Example 1 Increased Preservation of Food

A food packaging material is provided. The food packaging materialincludes a glass outer layer, a crosslinked polymer based matrix coatingthe inside of the outer layer, and an oxygen permeable polymer coatingthe top of the matrix. Within the matrix is a colony of Lactobacillusdelbrueckii subsp. Bulgaricus, and Bacillus subtilis as well as a foodsource for the colony. A piece of meat is placed within the foodpackaging material. Any oxygen present is converted to carbon dioxide bythe colony, thereby reducing the presence of oxygen and reducing theactivity of other microorganisms that depend upon oxygen for replicationand/or survival.

Example 2 Long Term Storage of Pharmaceuticals

A plastic packaging material is provided. The packaging materialincludes a rigid plastic outer layer, a starch based matrix coating theinside of the packaging material, and an oxygen permeable polymercoating on top of the matrix. Within the matrix is a colony ofNeurospora as well as a sugar based food source for the colony. Acollection of pills are placed within the plastic packaging material.Any oxygen present is converted by the colony to carbon dioxide, therebyreducing the presence of oxygen and reducing the oxidation of thepharmaceuticals.

Example 3 Method of Flavor Modification

A food packaging material is provided. The food packaging materialincludes a glass outer layer, a crosslinked polymer based matrix coatingthe inside of the packaging material, and an oxygen and lactic acidpermeable polymer coating on top of the matrix. Within the matrix is acolony of Streptococcus lactis. A quantity of milk is placed within thefood packaging material. Any oxygen present is converted by the colonyto carbon dioxide, thereby reducing the presence of oxygen and reducingoxidation of the milk. At the same time, the Streptococcus lactis canconvert the lactose in the milk to lactic acid, thereby creating buttermilk.

The present disclosure is not to be limited in terms of the particularembodiments described in this application, which are intended asillustrations of various aspects. Many modifications and variations canbe made without departing from its spirit and scope, as will be apparentto those skilled in the art. Functionally equivalent methods andapparatuses within the scope of the disclosure, in addition to thoseenumerated herein, will be apparent to those skilled in the art from theforegoing descriptions. Such modifications and variations are intendedto fall within the scope of the appended claims. The present disclosureis to be limited only by the terms of the appended claims, along withthe full scope of equivalents to which such claims are entitled. It isto be understood that this disclosure is not limited to particularmethods, reagents, compounds, compositions or biological systems, whichcan, of course, vary. It is also to be understood that the terminologyused herein is for the purpose of describing particular embodimentsonly, and is not intended to be limiting.

With respect to the use of substantially any plural and/or singularterms herein, those having skill in the art can translate from theplural to the singular and/or from the singular to the plural as isappropriate to the context and/or application. The varioussingular/plural permutations may be expressly set forth herein for sakeof clarity.

It will be understood by those within the art that, in general, termsused herein, and especially in the appended claims (e.g., bodies of theappended claims) are generally intended as “open” terms (e.g., the term“including” should be interpreted as “including but not limited to,” theterm “having” should be interpreted as “having at least,” the term“includes” should be interpreted as “includes but is not limited to,”etc.). It will be further understood by those within the art that if aspecific number of an introduced claim recitation is intended, such anintent will be explicitly recited in the claim, and in the absence ofsuch recitation no such intent is present. For example, as an aid tounderstanding, the following appended claims may contain usage of theintroductory phrases “at least one” and “one or more” to introduce claimrecitations. However, the use of such phrases should not be construed toimply that the introduction of a claim recitation by the indefinitearticles “a” or “an” limits any particular claim containing suchintroduced claim recitation to embodiments containing only one suchrecitation, even when the same claim includes the introductory phrases“one or more” or “at least one” and indefinite articles such as “a” or“an” (e.g., “a” and/or “an” should be interpreted to mean “at least one”or “one or more”); the same holds true for the use of definite articlesused to introduce claim recitations. In addition, even if a specificnumber of an introduced claim recitation is explicitly recited, thoseskilled in the art will recognize that such recitation should beinterpreted to mean at least the recited number (e.g., the barerecitation of “two recitations,” without other modifiers, means at leasttwo recitations, or two or more recitations). Furthermore, in thoseinstances where a convention analogous to “at least one of A, B, and C,etc.” is used, in general such a construction is intended in the senseone having skill in the art would understand the convention (e.g., “asystem having at least one of A, B, and C” would include but not belimited to systems that have A alone, B alone, C alone, A and Btogether, A and C together, B and C together, and/or A, B, and Ctogether, etc.). In those instances where a convention analogous to “atleast one of A, B, or C, etc.” is used, in general such a constructionis intended in the sense one having skill in the art would understandthe convention (e.g., “a system having at least one of A, B, or C” wouldinclude but not be limited to systems that have A alone, B alone, Calone, A and B together, A and C together, B and C together, and/or A,B, and C together, etc.). It will be further understood by those withinthe art that virtually any disjunctive word and/or phrase presenting twoor more alternative terms, whether in the description, claims, ordrawings, should be understood to contemplate the possibilities ofincluding one of the terms, either of the terms, or both terms. Forexample, the phrase “A or B” will be understood to include thepossibilities of “A” or “B” or “A and B.”

In addition, where features or aspects of the disclosure are describedin terms of Markush groups, those skilled in the art will recognize thatthe disclosure is also thereby described in terms of any individualmember or subgroup of members of the Markush group.

As will be understood by one skilled in the art, for any and allpurposes, such as in terms of providing a written description, allranges disclosed herein also encompass any and all possible subrangesand combinations of subranges thereof. Any listed range can be easilyrecognized as sufficiently describing and enabling the same range beingbroken down into at least equal halves, thirds, quarters, fifths,tenths, etc. As a non-limiting example, each range discussed herein canbe readily broken down into a lower third, middle third and upper third,etc. As will also be understood by one skilled in the art all languagesuch as “up to,” “at least,” and the like include the number recited andrefer to ranges which can be subsequently broken down into subranges asdiscussed above. Finally, as will be understood by one skilled in theart, a range includes each individual member. Thus, for example, a grouphaving 1-3 cells refers to groups having 1, 2, or 3 cells. Similarly, agroup having 1-5 cells refers to groups having 1, 2, 3, 4, or 5 cells,and so forth.

From the foregoing, it will be appreciated that various embodiments ofthe present disclosure have been described herein for purposes ofillustration, and that various modifications may be made withoutdeparting from the scope and spirit of the present disclosure.Accordingly, the various embodiments disclosed herein are not intendedto be limiting, with the true scope and spirit being indicated by thefollowing claims.

1. A packaging material comprising: an outer layer, wherein the outerlayer is effectively impermeable to oxygen; an inner layer, wherein theinner layer is effectively permeable to oxygen; and a bioactivematerial, wherein in the bioactive material is located between the innerlayer and the outer layer, wherein the bioactive layer comprises atleast one microorganism, and wherein the at least one microorganism issubstantially in a metabolically quiescent state.
 2. The packagingmaterial of claim 1, wherein the at least one microorganism comprises aliving microorganism.
 3. The packaging material of claim 1, wherein theat least one microorganism comprises a bacterium or a yeast.
 4. Thepackaging material of claim 1, wherein the at least one microorganism isaerobic.
 5. (canceled)
 6. (canceled)
 7. (canceled)
 8. The packagingmaterial of claim 1 further comprising a matrix positioned between theouter layer and the inner layer, wherein the at least one microorganismis associated with the matrix.
 9. The packaging material of claim 8,wherein the matrix comprises at least one of a starch, a carbohydrate, apolynucleotide, a protein, or a synthetic polymer.
 10. The packagingmaterial of claim 8, wherein the matrix comprises at least one of apeptide, a casein peptone, a vitamin, or a trace element.
 11. Thepackaging material of claim 1, wherein the inner layer comprises atleast one of a polymer film or an insert.
 12. The packaging material ofclaim 1, wherein the outer layer, inner layer, or both the outer layerand the inner layer provide a rigid wall of the packaging material. 13.The packaging material of claim 1, wherein the packaging material is inthe shape of a storage container.
 14. The packaging material of claim13, wherein the storage container is a food storage container. 15.(canceled)
 16. (canceled)
 17. (canceled)
 18. (canceled)
 19. Thepackaging material of claim 13, wherein the storage container is a bag.20. The packaging material of claim 19, wherein the bag is self sealing.21. A method of containing an item, the method comprising: providing apackaging material, the packaging material comprising: an outer layer,wherein the outer layer is effectively impermeable to oxygen; an innerlayer, wherein the inner layer is permeable to oxygen; and a bioactivematerial, wherein in the bioactive material is located between the innerlayer and the outer layer, and wherein the bioactive layer comprises atleast one microorganism, wherein the at least one microorganism issubstantially in a metabolically quiescent state; and exposing an itemto the packaging material, wherein a surface of the inner layer faces asurface of the item, thereby containing the item.
 22. The method ofclaim 21, wherein the inner layer is proximal to the item and the outerlayer is distal to the item.
 23. The method of claim 21, wherein theitem comprises a perishable food product.
 24. The method of claim 21,wherein the packaging material forms at least a part of a container. 25.The method of claim 24, wherein exposing the item to the packagingmaterial comprises placing the item within the container.
 26. (canceled)27. (canceled)
 28. The method of claim 25, further comprising sealingthe container with the item inside of the container to provide a sealedcontainer, wherein when the item is within the sealed container, thereis a headspace volume.
 29. The method of claim 28, wherein the headspacevolume comprises a first percent of oxygen immediately after thecontainer is sealed.
 30. The method of claim 29, wherein the headspacevolume comprises a second percent of oxygen 2 hours after the containeris sealed, wherein the second percent is less than the first percent.31. (canceled)
 32. The method of claim 28, wherein an amount of oxygenwithin the headspace volume passes through the inner layer to thebioactive layer, and wherein the amount of oxygen is exchanged with anamount of carbon dioxide by the at least one microorganism.
 33. A methodof making a packaging material, the method comprising: providing anouter layer, wherein the outer layer is substantially impermeable tooxygen; providing a bioactive material, wherein providing a bioactivematerial comprises: providing at least one microorganism; and inducingthe at least one microorganism into a metabolically quiescent state; andproviding an inner layer, wherein the inner layer is permeable tooxygen, and wherein the bioactive material is positioned between theouter layer and the inner layer, thereby making a packaging material.34. The method of claim 33, wherein providing a bioactive materialfurther comprises: embedding the at least one microorganism in a matrix,thereby providing the bioactive material.
 35. The method of claim 34,wherein embedding the at least one microorganism comprises combining theat least one microorganism and a matrix material and cross-linking,polymerizing, or both cross-linking and polymerizing the matrixmaterial.
 36. (canceled)
 37. (canceled)